xMAP® Connect: Predicting Treatment Response in Breast Cancer

MD Anderson pathologist shares results from xMAP®-powered breast cancer studies at user group meeting

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The recent xMAP® Connect user group meeting featured an impressive roster of speakers, and included among them was Fraser Symmans, pathologist and director at MD Anderson Cancer Center. His presentation offered an update on those projects designed to predict an individual’s breast cancer response to endocrine therapy—which is one of the most important and widely used long-term treatments available for those with HR+/HER2- breast cancer. Moreover, those clinical assays that interrogate therapeutic response have been supported by Luminex’s xMAP Technology.

xMAP Technology helps build platform for breast cancer studies

Symmans explained that his work began years ago, when his earliest assays were designed from microarrays that enabled the analysis of key transcripts. But as his team progressed, they aimed to create a customized assay that might eventually become a clinical test. They began by establishing a set of genes whose expression revealed information about the activity of an individual’s endocrine pathway and their correlation with cancer treatment. Over time, the scientists gathered data on endocrine therapy sensitivity, response to chemotherapy, burden of disease at diagnosis, and more.

Ultimately, Symmans’s team selected a group of genes that collectively serve as a prognostic index for endocrine therapy response. However, they wanted to move forward with an assay platform that could be used with Formalin-Fixed Paraffin-Embedded (FFPE) samples. Symmans and his colleagues compared an xMAP assay to the original microarrays, as well as to another gene expression technology and found excellent concordance across the platforms.

xMAP Technology serves as “a very stable platform”

“We made the choice for xMAP based on some preferences and some workflow logistics that we thought would be advantageous,” he said. It offers “outstanding performance” and now serves as “a very stable platform for us,” he added. Subsequent inter- and intra-lab studies demonstrated the platform’s strong reproducibility and “excellent concordance of this assay in different labs,” Symmans reported — noting that this was the case even for users who had no previous familiarity with the technology.

Symmans and his colleagues have used the xMAP-based assay to study various types of breast cancer samples as they expand their validation of the prognostic index. They have now assessed clinical utility in several different projects, predicting not only response to endocrine therapy, but also long-term survival. Their assay is now established in a CLIA laboratory, ready for use in prospective clinical trials and/or in commercial development projects.

Learn more about how xMAP Technology can be used for a wide variety of applications.

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xMAP® Connect: Measuring Immune Response for Vaccine Development

Texas Children’s Hospital scientist speaks at our recent user group meeting about low-cost COVID‑19 vaccine

At the recent xMAP® Connect user group meeting, we were honored to host Jeroen Pollet, assistant professor at Baylor College of Medicine, who also serves as a director on the Vaccine Development Team at Texas Children’s Hospital. Pollet presented his insights regarding the application of xMAP Technology towards a multiplex cytokine release assay that measures immune response to vaccine candidates for tropical and other diseases.

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Typically, Pollet and his team focus on surveillance, diagnostics, and vaccine development for neglected tropical diseases. “What’s truly unique about us is our science,” he said. The Vaccine Development Center is part of a special program that combines basic research with product development, as it spans preclinical and clinical testing—even seeing it through the regulatory review process. “It’s an integrated product development strategy,” Pollet added.

Shifting gears for COVID‑19

Upon the emergence of COVID-19, Pollet’s team turned its attention to this new public health threat and developed Corbevax®, which is an effective, low-cost vaccine that could supply low-income countries where other COVID-19 vaccines are cost-prohibitive. In addition to having received emergency use authorization and being commercially available, Corbevax has been licensed for production in India, where 300 million doses are being manufactured. There, it has already been used to vaccinate more than 10 million children aged 12 to 14. Moreover, 100 million vaccine doses have been approved and designated for Botswana, where manufacturers will begin ramping up local production.

Multiplexing technology offers ease of use, standardization, and the ability to query multiple analytes

This exciting development was made possible through the dedicated and tireless work of Pollet and his colleagues, who relied on Luminex’s xMAP Technology to make it happen. They used Luminex’s bead-based multiplexing platform in a pipeline with orthogonal technologies to test for an immune response following immunization. They chose the multiplex cytokine release assay for its ease of use, standardization, and its ability to query multiple analytes, Pollet informed. In preclinical studies involving mice, the assay allowed them to quantify the cytokines secreted after two doses of the vaccine. Subsequent studies were also performed in primate models. “It’s really important that we get data as fast as possible,” Pollet said. Because xMAP allows testing for multiple analytes at once, time to results is accelerated by eliminating the need for tedious serial testing.

xMAP Technology supports vaccine development

Pollet’s team works with both recombinant protein- and mRNA-based vaccines. In separate studies involving Chagas disease, Pollet relayed that a heterologous approach using both protein and mRNA looked very promising towards boosting immune response based on cytokine assay results. “We believe that heterologous vaccination is really the way forward,” he said.

Learn more about how xMAP Technology can be used to support vaccine development.

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Multiplexing 101: New Resource for Immunoassays in Biomarker Discovery

Educational eBook offers tips and helpful examples

If you work with biomarker discovery, don’t miss out on this free eBook exploring how scientists can utilize multiplexing immunoassay technology in their research to generate more answers in less time with better reproducibility. Multiplex technology provides a high-throughput approach to biomarker research that saves time and money, minimizing the amount of samples and reagents required to perform an immunoassay.

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Produced by BioCompare®, this eBook offers a variety of helpful tips for using multiplex tools as well as key tutorials, case studies, and other resources.

Some of the eBook highlights include:

  • An introductory guide comparing multiplex immunoassays to single-plex immunoassays
  • A look at how multiplex assays aid in the interrogation of conditions related to cytokines
  • A case study demonstrating how multiplex assays can be used to detect protein and oligo targets by deploying a dual-reporter feature
  • A comparison of multiplex assays to electrochemiluminescence showing differences in throughput and reliability of detection
  • A look at simplified data analysis using improved immunoassay software

Backed by more than 50,000 publications, multiplexing immunoassay technology is a proven alternative to traditional ELISAs that can help scientists scale up their biomarker discovery operations while preserving samples.

For more information on multiplexing for biomarker discovery, download this free ebook today.

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CDC Scientist Describes Multiplex Testing for Malaria at Immunology2022™

Antigen and antibody tests generate important data for characterizing population-level trends, individual immune response, and more

At Immunology2022™, this year’s meeting of the American Association of Immunologists, Eric Rogier, a microbiologist from the CDC, offered an exciting presentation about utilizing bead-based multiplex assays for generating epidemiological and immunological data about malaria.

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Dr. Rogier, a specialist in malaria and other tropical diseases, aims to develop reliable diagnostic and serological tests to aid public health experts in their understanding of outbreaks, disease trends, and immune response. Unlike many tropical diseases that are easy to identify, malaria causes non-specific symptoms (such as fever, chills, headache, and nausea) that can be difficult to diagnose. In addition, asymptomatic infections are common in areas with high rates of transmission. This makes it difficult to estimate the true burden of disease, he noted.

Searching for several antigens at once with xMAP® Technology

Obtaining an accurate case count would allow researchers to track the effectiveness of treatments, monitor progress over time, and perhaps ultimately achieve disease elimination, Rogier informed. Thus, researchers require epidemiological data that is both accurate and plentiful. Unfortunately, the data generated by local health facilities is not always reliable or accessible.

To address these challenges, Rogier and his colleagues have developed multiple tests based on

His team has created several xMAP assays for malaria, with varying numbers of analytes. They have used a 9-plex assay to capture antigens of interest, including a Nigerian household survey that tested samples collected from more than 31,000 children. A separate study in Djibouti averaged 1,000 people and even allowed scientists to identify malaria co-infections of P. falciparum and P. vivax, thereby highlighting xMAP Technology’s cost advantage. Rather than spending $10 per sample on PCR tests for each person, the team spent $2 per sample and the xMAP assay yielded the same data, with results more comprehensive than any single-target test.

“Antibodies don’t lie”

Eric Rogier, PhD MPH, Microbiologist at the Center for Disease Control & Prevention (CDC)

Eric Rogier, PhD MPH, a microbiologist at the CDC, and his team have created several xMAP assays for malaria, with varying numbers of analytes.

In addition, Rogier has developed serological assays for antibody detection. “You’re looking at a history of exposure for an individual versus an active infection,” he said. This approach provides “an objective measure of malaria exposure in [a] population,” he added. “Antibodies don’t lie.”

Serology testing is especially useful in low-transmission settings, where generating enough data for epidemiological use can be challenging. In one example, Rogier showed that the antibody test identified 50 times more positive samples than tests for active infections. In another, he noted that serology data “can be a very powerful metric to assess population-level exposure.” Thanks to multiplexing, the tests can measure IgM, IgA, IgE, and all four subclasses of IgG.

Generating data for malaria—multiplexing makes it possible

Furthermore, a longitudinal study conducted in Angola offered scientists a view of the immune response to malaria, wherein patients were followed for 42 days with weekly sampling and serological testing. This study involved a 37-plex xMAP assay and led to new insights into antibody half-life and other dynamics. Interestingly, the results also made it possible to distinguish between patients who had malaria for the first time and those who had been infected more than once.

Together, these studies demonstrate that xMAP-based assays can cost-effectively generate rich epidemiological data for malaria in a variety of settings.

What range of applications can xMAP® Technology help with?

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Luminex Licensed Technologies Partner Spotlight: VRL

For streamlined animal health testing with robust results, VRL chooses xMAP® Technology

Diagnostic testing isn’t just important for human health—it’s a key component of maintaining animal health as well. Few people are as familiar with this as the scientific experts at VRL, a diagnostic laboratory that has been serving animal health needs for more than 30 years.

VRL specializes in monitoring the health of animals used in research studies, though it also performs services for veterinarians in other areas of animal health. In its laboratory facilities, VRL scientists aim to detect viruses, bacteria, parasites, and other pathogens that could make animals sick.

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Researchers at VRL use extensive diagnostic testing to ensure that animals—primarily rodents and nonhuman primates—are healthy prior to any research study. Their goal is to minimize variables that might confound study results. For example, a mouse that appears healthy, but covertly harbors a pathogen affecting its immune system could respond very differently than expected within an experiment focused on immune-related results.

Saving time and precious sample

Because VRL labs receive thousands of samples each day, their work requires a high-throughput approach with reliable results. Arlene Leon, director of operations and scientific affairs at VRL, is responsible for ensuring that all testing is performed properly, on time, and maintains standards of excellent scientific quality.

Under her leadership, VRL has implemented Luminex Licensed Technologies Partner, receiving early access to new kits and products as well as enhanced support.

“Luminex takes us to the next level. We get more statistical data and more reliable results.”

Arlene Leon, director of operations and scientific affairs at VRL
— Arlene Leon
Director of Operations and Scientific Affairs at VRL

The need for a multiplexed approach was obvious to Leon. Clients may ask for as many as 23 different tests on a single animal, and there is rarely enough sample available to perform these tests as individual ELISAs. Adopting xMAP’s bead-based multiplexing technology has allowed her team to overcome this challenge. “With Luminex, we use very little sample volume, and we can put all 23 tests in a single well,” Leon says. xMAP’s multiplexed approach also minimizes hands-on time for technicians, makes it possible to run more samples at once, and generates answers more quickly.

“Luminex takes us to the next level,” Leon says. “We get more statistical data and more reliable results.” The approach also allows her team to continue meeting their desired turnaround time of 24 to 48 hours, even as more samples come in.

Panel-based testing to xMAP multiplexing

Leon’s team is in the process of translating their panel-based testing approach to xMAP assays. xMAP’s multiplexed assays help optimize the team’s work, as each type of animal requires testing for different pathogens. Nonhuman primates, for example, are typically tested for five pathogens, while mice may need testing for more than two dozen pathogens. VRL aims to begin returning results for its first panel in the coming months, and later, transfer more of their clients to xMAP assays.

As xMAP Technology is rolled out in the VRL labs, Leon says it will also make it possible for VRL to offer new services, such as biomarker detection. “Now, the sky’s the limit in what our clients can ask us to do for them,” she says.

“As a high-throughput [veterinary] diagnostic lab with very short turnaround times, we have to make sure we have high quality and excellent client services,” Leon adds. “Working with Luminex helps us to provide that level of service.”

VRL is one of our exclusive partners that span a wide range of industries and specialties. See our website to view a full list of our partners and click here to learn more about partnering with Luminex.

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Webinar: How Drug Discovery Scientists Improved Biomarker Evaluation with xMAP® Technology

To better understand a candidate therapy for neurodegenerative disorders, researchers needed an easy-to-use multiplexing technology

At the Labroots-hosted Neuroscience 2022 meeting, Diana Price from Neuropore Therapies offered a fascinating presentation about how her team is using xMAP® Technology to characterize candidate therapies in order to treat Parkinson’s disease, amyotrophic lateral sclerosis (ALS), and other neurodegenerative disorders. The one-hour talk is now available on-demand, with the option to earn P.A.C.E. CE and Florida CE credits.

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Neuropore Therapies is a San Diego-based biopharma company that develops small-molecule therapies to treat neurological diseases. Their scientists aim to address the core mechanisms that underly these health conditions, in the hopes that a viable treatment can be used for more than one disorder. Among the most common disease features are inflammation in the brain and/or spinal cord.

Implementing a development strategy using xMAP Technology

Diana L. Price, PhD, Executive Director, Biosciences, Neuropore Therapies, Inc.

Diana L. Price, PhD
Executive Director, Neuropore Therapies, Inc.

Price noted that most neurodegenerative diseases lack validated clinical biomarkers, which presents a challenge for both preclinical and clinical development. To address this, her team has implemented a translational biomarker development strategy that incorporates measurements from many different biological signals. This approach could influence not only when confirming treatment effects, but also aid in identifying biomarkers that may prove useful for future clinical trials. Thus, these biomarkers could serve as inclusion or exclusion criteria as they help to manage variables within the patient population.

For this strategy, Price and her team searched for technologies that could generate multiplexed, multimodal, and unbiased data in support of their development programs. The ideal tools would be flexible, efficient, and versatile. They selected xMAP Technology, which allowed them to measure hundreds of expressed genes or proteins at a time. According to Price, her team now deploys this technique as a “key technology” when characterizing, developing, and validating in vivo models, as well as when profiling in vivo target engagement.

Using multiplexing technology for the study of neurodegenerative disorders

Luminex Dot Pattern

To demonstrate how her team uses this multiplexing technology, Price shared details about candidate therapies that function as antagonists for toll-like receptors 2 and 9 (TLR2 and TLR9), which are the proteins associated with Parkinson’s disease and other neurodegenerative disorders. These proteins trigger a cellular inflammatory response based on the presence of pathogens or damage, which are the cause behind these neurodegenerative disorders.

An evaluation of more than 62,000 compounds revealed some candidates with promising potency and specificity for TLR2 and TLR9. Her team used xMAP Technology to perform multiplexed evaluations of plasma cytokines in wild type mice to better understand these candidate molecules.

Currently, they are evaluating these candidates for Parkinson’s disease, ALS, and Guillain-Barré syndrome, all of which are highly associated with TLR2 and TLR9. In the preclinical animal models for each disorder, it was shown that the candidate small-molecule improves multiple traits, and thus reduces protein pathology and inflammatory markers, while boosting overall health.

Learning more about xMAP Technology

xMAP Technology has made it simple for the researchers to evaluate the many different TLR and inflammation markers, as they seek to characterize the animal samples’ response to treatment. “It was such an easy evaluation to run,” Price said. Her team is very happy with how the animal models are responding. Meanwhile, some of the study results thus far have also highlighted certain biomarkers that may prove useful for measuring treatment response and/or for clinical trial criteria.

Learn more about how xMAP Technology could be useful for your research.

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Peer-Reviewed Publications Illustrate Versatility of xMAP® Technology

Applications go well beyond human samples, with demonstrated uses in animals, bacteria, and more

When the Luminex team first conceived the idea of the bead-based multiplexing that would lead to xMAP® Technology, they never imagined the broad range of applications it would one day enable. Decades later, we are constantly impressed by the creative uses scientists find for this versatile platform.

To celebrate the great diversity of xMAP-enabled applications in the research community, we’ve pulled together a sampling of recent publications showing how this technology is being used beyond human health needs. From animals to viruses and more, here’s a look at the utility of bead-based multiplexing in a variety of applications.

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Evaluation of serum MMP-2 and MMP-3, synovial fluid IL-8, MCP-1, and KC concentrations as biomarkers of stifle osteoarthritis associated with naturally occurring cranial cruciate ligament rupture in dogs

Journal: PLoS One

First author: Sarah Malek

Project summary: Scientists aimed to identify a biomarker signature associated with inflammation in dogs with stifle osteoarthritis, a progressive degenerative canine disease.

How xMAP made a difference: xMAP Technology was used to analyze serum and synovial fluid samples collected from 62 affected and 50 healthy dogs. The team found a biomarker signature strongly linked to inflammation among dogs with the disease.

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Detection of specific Atlantic salmon antibodies against salmonid alphavirus using a bead-based immunoassay

Journal: Fish & Shellfish Immunology

First author: Lena Hammerlund

Project summary: Scientists aimed to develop a reliable serological assay to detect antibodies indicative of salmonid alphavirus, a cause of pancreas disease in Atlantic salmon and a major threat for the aquaculture industry in Europe.

How xMAP made a difference: xMAP Technology enabled scientists to design a bead-based immunoassay that takes less time than a virus neutralization assay and can be used on live fish due to the small sample volume required. Results were highly correlated with those from the neutralization assay.

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Detection of Babesia, Borrelia, Anaplasma, and Rickettsia spp. in Adult Black-Legged Ticks (Ixodes scapularis) from Pennsylvania, United States, with a Luminex Multiplex Bead Assay

Journal: Vector-Borne and Zoonotic Diseases

First author: Julia Livengood

Project summary: Scientists aimed to detect the presence of a range of microorganisms harbored by the black-legged tick, Ixodes scapularis, in a Pennsylvania study.

How xMAP made a difference: The team used an xMAP-based assay to identify 20 pathogens in an analysis of nearly 300 tick samples.

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Establishment and application of a 10-plex liquid bead array for the simultaneous rapid detection of animal species

Journal: Journal of the Science of Food and Agriculture

First author: Mingzhu Mei

Project summary: Researchers aimed to address the growing challenge of meat fraud and adulteration, problems that are on the rise in regions around the world.

How xMAP made a difference: The research team developed a multiplex xMAP assay to detect 10 different types of animals in just three hours, and then validated the assay by testing it on DNA extracts from meat products as well as animal feed. Results showed excellent concordance with conventional testing methods.

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Bio-Plex suspension array immuno-detection of Listeria monocytogenes from cantaloupe and packaged salad using virulence protein inducing activated charcoal enrichment media

Journal: Food Microbiology

First author: J.B. Day

Project summary: Scientists aimed to improve the testing process for detecting the Listeria monocytogenes pathogen in food samples, such as salad greens and cantaloupe.

How xMAP made a difference: Scientists selected a different type of enrichment media and deployed an xMAP-based assay to detect even small traces of the pathogen, with results generated in one day.

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Establishment and optimization of a liquid bead array for the simultaneous detection of ten insect-borne pathogens

Journal: Parasites & Vectors

First author: Hui-yu Wang

Project summary: Insect-borne pathogens are a major public health issue, with notorious examples such as Ebola virus and West Nile fever virus. In this study, scientists focused on developing a multiplex assay to detect 10 of these pathogens.

How xMAP made a difference: Researchers chose xMAP as their platform, creating a 10-plex assay and testing 3,000 clinical samples for validation. Detection of all pathogens was highly sensitive, specific, and reproducible, with positive results seen for as few as 10 copies of the target gene.

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Learn more about xMAP Technology and how it could make a difference in your lab.

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    • Multiplexing Technology Supports Autoantibody Studies for Immuno‑Oncology Treatment

    Scientists use xMAP® Technology to detect thousands of antibodies in hundreds of samples

    At the recent Drug Discovery and Development 2022 conference, Oncimmune® Chief Technical Officer, Hans-Dieter Zucht, gave a presentation about how his company’s scientists are using autoantibodies as biomarkers for a broad range of immuno-oncology studies. This work is made possible with bead-based multiplexing

    Zucht pointed out that while immunotherapies have been remarkably effective in fighting cancer, they only work for a small fraction of patients with cancer. Those who do respond well may also suffer from immune-related adverse events to the treatment. There is a pressing need to identify biomarkers that may help stratify patients to ensure immunotherapies are directed to the people most likely to benefit, as well as biomarkers that could predict the likelihood of adverse events.

    Insight into autoantibodies

    “The concordance and repeatability of these measurements is very high.”

    Hans-Dieter Zucht, Chief Technical Officer at Oncimmune
    — Hans-Dieter Zucht
    Chief Technical Officer at Oncimmune

    At Oncimmune, scientists have turned their attention to autoantibodies. This class of antibodies has already been used to elucidate clinically relevant interactions between tumor cells and the immune system. Oncimmune offers the check out this study of patients with metastatic urothelial carcinoma or see our

    Learn more about Oncimmune’s approach to multiplexed autoantibody pofiling of patients with metastatic urothelial carcinoma.

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